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作物学报 ›› 2012, Vol. 38 ›› Issue (07): 1232-1239.doi: 10.3724/SP.J.1006.2012.01232

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

烟草细胞质雄性不育系及其保持系的花蕾差异蛋白质分析

祁建民1,**，马红勃1,2,**，徐建堂1，陈美霞1,3，周东新4，王涛5，陈顺辉6   

  1. 1 福建农林大学生命科学学院 / 作物遗传育种与综合利用教育部重点实验室, 福建福州 350002; 2 南京农业大学作物遗传与种质创新国家重点实验室, 江苏南京 210095; 3 宁德师范学院生物工程系, 福建宁德 352100; 4 福建省龙岩市烟草科学研究所, 福建龙岩 364000; 5 福建省南平市烟草公司, 福建南平 353200; 6 福建省福州市烟草科学研究所, 福建福州 350003
  • 收稿日期:2011-11-17 修回日期:2012-02-22 出版日期:2012-07-12 网络出版日期:2012-04-06
  • 基金资助:

    本研究由中国烟草总公司重点项目(110201002006)资助

Proteomic Analysis of Bud Differentiation between Cytoplasmic Male-Sterile Line and Maintainer in Tobacco

QI Jian-Min1,**,MA Hong-Bo1,2,**,XU Jian-Tang1,CHEN Mei-Xia1,3,ZHOU Dong-Xin4,WANG Tao5,CHEN Shun-Hui6   

  1. 1 Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2 National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China; 3 Ningde Normal University Biological Engineering, Ningde 352100, China; 4 Longyan Tobacco Branch Company, Longyan 364000, China; 5 Nanping Tobacco Company, Nanping 353200 China; 6 Fuzhou Institute of Tobacco Science of Fujian Province, Fuzhou 350003, China?
  • Received:2011-11-17 Revised:2012-02-22 Published:2012-07-12 Published online:2012-04-06

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1475

被引次数

12

摘要： 为探讨烟草雄性不育的分子遗传机理，对烟草细胞质雄性不育系和保持系进行差异蛋白质组学研究。采用固相pH梯度SDS-PAGE双向电泳，对烟草细胞质雄性不育系MSK326及其保持系K326雄蕊原基分化期、四分体时期及单核时期花蕾蛋白质进行分离，经考马斯亮蓝染色，获得了分辨率和重复性较好的双向电泳图谱。使用PDQuest软件分析蛋白质图谱，利用基质辅助激光解吸电离飞行时间串联质谱分析，然后用Mascot软件对NCBInr数据库搜索。在分子量14.4~97.6 kD、等电点4~7线性范围内，检测到约365个蛋白点，其中7个蛋白质点在保持系中出现, 在不育系中缺失，分别被鉴定为核酮糖-1,5-二磷酸羧化酶/加氧酶、多酚氧化酶、Patatin homolog蛋白、PSI 9 kD蛋白4类蛋白，推测不育系MSK326雄性不育性可能与营养代谢紊乱、间接抑制雄性器官发育、养分供给不足、免疫能力低和能量转移受到抑制等有关。

关键词: 烟草, 细胞质雄性不育, 花蕾, 差异蛋白质, 双向凝胶电泳

Abstract: To explorethe moleculargenetic mechanismsof cytoplasmic male sterility in tobacco,we studied the differential protein between the cytoplasmic male-sterile line and its maintainer. Immobilized pH gradient two-dimensional gel electrophoresis (2-DE) technique was used to separate the protein spots while gels were stained with the Coomassie Blue G-250. The difference between protein maps of bud from cytoplasmic male-sterile line MSK326 and maintainer line K326 was analyzed with PDQuest image software. Matrix-assisted laser-adsorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) technique was used to obtain the peptide mass of differentially expressed protein spots. The Mascot software was used to search the protein database NCBInr to identify those spots interested. A total of 365 protein spots were detected within Mr 14.4–97.6 kD and pH 4–7. Seven protein spots appeared in the protein map of maintainer line K326 but absent in that of CMS line MSK326, which were identified as ribulose-1,5-bisphosphate carboxylase/oxygenase, polyphenol oxidase, patatin homolog, PSI 9 kD protein. It was inferred that the male sterility of MSK326 might be related to energy metabolism turbulence, abnormality in male organ development, deficient nutrition supply and immunity, and inhibition of energy transfer.

Key words: Tobacco, Cytoplasmic male sterility, Bud, Differential protein, 2-DE

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